We aim to develop a fully integrated microfluidic immunodiagnostic platform utilizing a novel air-liquid cavity acoustic transducer (ALCAT) technology pioneered in the lab of Dr Abe Lee. The diagnosis platform is based on the well-established clinical diagnostic tool, the enzyme immunoassay (eg ELISA). ELISAs are an integral platform for clinical diagnosis because they are rapid, robust, and provide results that are highly sensitive and specific. Additionally, ELISAs rely on the inherent humoral immune response of the host to produce antibodies indicative of pathological status. In this proposal, we will form a strategic partnership of expertise in protein microarrays (Antigen Discovery, Inc. and Dr Phil Felgner) and in microfluidics (Dr Abe Lee) to develop a point-of-care diagnostic system consisting of a cost-effective disposable microfluidic plastic chip and a portable analyzer that can be driven by a mobile computing device for rapid serological evaluation of Melioidosis. By synergistically incorporating novel ALCAT-based approaches into these arrays we will have the versatility and power to reduce enzymatic development times, limit production costs, and fully integrate all components into a disposable assay. From previous NIH/NIAID funding we have discovered serodiagnostic antigens for 12 NIAID priority agents (Brucella melitensis, Francisella tularensis, Burkholderia pseudomallei, Coxiella burnetii, Salmonella enterica typhi, Rickettsia rickettsii, Orientia tsutsugamushi, Toxoplasma gondii, M. tuberculosis, variola major related pox viruses, Dengue, and West Nile). We have also identified serodiagnostic antigens for non-biodefense targets including: Bartonella henselae, Borrelia burgdorferi, Hepatitis B Virus, Chlamydia, Plasmodium falciparum, Human papillomavirus, and Herpes simplex virus. This approach will leverage our existing content of serodiagnostic antigens (and appropriate banked sera collection) to produce a multiplex device that will contain multiple antigens from numerous pathogens and enable clinicians to determine which pathogen an individual was exposed to. We believe the platform takes advantage of this wealth of content, and marries it to a versatile ALCAT-based microfluidics approach to produce a true lab-on-a- chip tool that is equipment free, technical expertise unrestricted, and useful for both developed and undeveloped countries. This approach produces a an immensely powerful diagnostic platform that will compliment current assays, provide diagnosis for a broad range of diseases, and is capable of utilizing additional enzyme immunoassay-based content. This proposal is an effective integration of emerging microarray technologies and will set the stage for future serological detection systems. PUBLIC HEALTH RELEVANCE: Centralized testing in well-resourced reference laboratories cannot adequately address the unpredictable demand for rapid diagnostics during mass disasters, natural disease outbreaks, and bioterrorist threats. During such emergencies, first responders must rely on accurate diagnostics that are quickly and efficiently deployable to patients who may not have easily recognizable signs or symptoms of an infection. Point-of-care (POC) multiplex diagnostics are well-suited to fill this important role as they are highly portable do not require extensive training, and provide prompt sample-to-answer times.